Capillary liquid cooling column temperature box and capillary electrophoresis analyzer

Abstract

The present invention is specifically a capillary liquid-cooled column temperature box and a capillary electrophoresis analyzer, the capillary liquid-cooled column temperature box comprises: a box body; a temperature regulating container and a liquid storage container arranged in the box body, a temperature regulating device is provided on one side of the temperature regulating container to fit with it, the temperature regulating container and the liquid storage container form a first circulation loop through a circulation pump and a circulation pipeline; the liquid storage container forms a second circulation loop through a capillary and a peristaltic pump, the second circulation loop comprises a first capillary, a second capillary and a third capillary, one end of the first capillary is connected to the first liquid outlet of the liquid storage container, one end of the third capillary is connected to the first liquid inlet of the liquid storage container, the other ends of the first capillary and the third capillary are respectively connected to the two ends of the second capillary through a tee; the tee is used to puncture the capillary, the capillary penetrates from one end of the first tee, passes through the second capillary, and penetrates from one end of the second tee. The present invention has the following advantages: providing continuous and stable heat exchange cooling for the capillary; optimizing the temperature stability during the capillary cooling process.

Description

Technical Field

[0001] The invention relates to the technical field of liquid phase analysis instruments, in particular to a capillary liquid cooling column temperature box and a capillary electrophoresis analyzer. Background Art

[0002] Capillary electrophoresis (CE) is an efficient separation and analysis technique that separates charged particles based on the difference in migration speed in the capillary under the action of an electric field. This technology rose rapidly in the late 1980s and has been widely used in the separation and analysis of small molecules, small ions, peptides and proteins due to its advantages such as rapidity, high efficiency, high sensitivity, easy quantification, good reproducibility and automation. Capillary electrophoresis technology separates in an extremely fine capillary column, increases the electric field strength to 100-200V / cm, and comprehensively improves the separation quality. Capillary electrophoresis includes a variety of modes, such as capillary zone electrophoresis (CZE), micellar electrokinetic capillary chromatography (MECC), capillary gel electrophoresis (CGE), etc., which can adapt to different separation needs.

[0003] In the prior art, during the process of separating liquid samples by capillary electrophoresis, excessive current will cause Joule heating, and the continuous accumulation of this heat is prone to produce bubbles. The formation of bubbles will interfere with the current stability during the electrophoresis process, affecting the migration speed and separation efficiency of the sample. The Joule heating effect will change the viscosity of the solution, thereby changing the time it takes for the sample zone to reach the detection window, affecting the separation effect. Controlling Joule heat is crucial to maintaining the separation and analysis effect of capillary electrophoresis. Therefore, a capillary liquid-cooled column oven and a capillary electrophoresis analyzer are now designed to continuously and evenly dissipate heat from the capillary without interfering with the normal capillary electrophoresis operation. Summary of the invention

[0004] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a capillary liquid-cooled column oven and a capillary electrophoresis analyzer to achieve continuous and uniform heat dissipation of the capillary during operation, and further control the heat exchange process to remain moderate to avoid condensation caused by sudden heat exchange in the storage line.

[0005] In order to achieve the above-mentioned purpose, a capillary liquid-cooled column temperature box is designed, comprising a box body, and also comprising: a temperature regulating container and a liquid storage container arranged in the box body, a temperature regulating device is provided on one side of the temperature regulating container to fit with the temperature regulating container, and the temperature regulating container and the liquid storage container form a first circulation loop through a circulation pump and a circulation pipeline; the liquid storage container forms a second circulation loop through a capillary and a peristaltic pump, and the second circulation loop comprises a first capillary, a second capillary and a third capillary, one end of the first capillary is connected to a first liquid outlet of the liquid storage container, one end of the third capillary is connected to a first liquid inlet of the liquid storage container, and the other ends of the first capillary and the third capillary are respectively connected to two ends of the second capillary through a tee; the tee is used to puncture the capillary, so that the capillary penetrates from one end of the first tee, passes through the second capillary, and then penetrates from one end of the second tee.

[0006] Preferably, the present invention also includes: an opening and closing cover is provided on the front side of the box body, one end of the opening and closing cover is pin-connected to the front side surface of the box body, the opening and closing cover and the front side surface of the box body form a cavity structure, and the capillary tube of the second circulation loop passes through the through hole on the front side surface of the box body and is arranged in the cavity structure.

[0007] Preferably, the present invention further comprises: the temperature regulating device comprises a plurality of heat dissipation fins and a plurality of fans, and the heat dissipation fins and the fans are arranged alternately.

[0008] Preferably, the present invention also includes: the temperature regulating container includes a main circulation pipe and a plurality of metal branch circulation pipes, the branch circulation pipes are closed and connected with the main circulation pipe after passing through the temperature regulating device, the main circulation pipe is also closed and connected with the first circulation loop, and the temperature regulating container is wrapped with insulation material on the outside of the main circulation pipe and the branch circulation pipe.

[0009] Preferably, the present invention further comprises: a PCB board is further provided in the box body, and the PCB board is in signal communication with the temperature regulating device, the circulation pump of the first circulation loop and the peristaltic pump of the second circulation loop to form signal control.

[0010] Preferably, the present invention further comprises: the capillary entry end of the first tee and the capillary exit end of the second tee are provided with disposable piercing rubber heads, so that the capillary pierces through and forms a liquid seal between the capillary and the tee through elastic deformation.

[0011] Preferably, the present invention also includes: characterized in that the flow rate of the peristaltic pump is 0.5 ml / s.

[0012] The present invention also provides a capillary electrophoresis analyzer, comprising: a capillary, both ends of the capillary are respectively immersed in a buffer solution; a sample injection head arranged on the capillary; an electrophoresis zone arranged on the capillary and on the right side of the sample injection head; a detector arranged on the capillary and on the right side of the electrophoresis zone; two buffer tanks arranged on both ends of the capillary and used to accommodate buffer solutions; a high-voltage electrical system whose both ends are electrically conductive with the buffer solutions in the two buffer tanks, and the portion of the capillary between the sample injection head and the detector is arranged in the second capillary of any of the above-mentioned capillary liquid-cooled column temperature boxes.

[0013] Compared with the prior art, the present invention has the following advantages:

[0014] By designing a multi-section circulation loop, the heat exchange liquid on the outside of the capillary tube always keeps flowing, providing continuous and stable heat exchange cooling for the capillary tube; through the structural design of the capillary tube and the tee, the capillary tube can obtain heat exchange during the working process, and the heat exchange liquid will not affect the liquid to be analyzed in the capillary tube; through the peristaltic pump and capillary tube design, the heat conduction effect of the heat exchange liquid on the capillary tube is more moderate, avoiding the condensation effect caused by sudden temperature changes; through the design of the opening and closing cover, capillary tube and box body, the capillary tube is isolated from most of the heat exchange effects with room temperature during the heat exchange process in the capillary tube, further optimizing and stabilizing the temperature stability during the capillary tube cooling process. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 , is a schematic diagram of the structure of the present invention;

[0016] Figure 2 , is a front view of the present invention;

[0017] Figure 3 ,yes Figure 2 AA plane section view;

[0018] Figure 4 ,yes Figure 2 The perspective view of the opening and closing cover is hidden;

[0019] Figure 5 , is a chromatogram in Example 1 without using the capillary liquid-cooled column oven of the present invention;

[0020] Figure 6 , is a chromatogram of the capillary liquid-cooled column oven of the present invention in Example 1;

[0021] In the figure: 1 box body, 2 temperature regulating container, 2.1 main circulation pipe, 2.2 branch circulation pipe, 3 liquid storage container, 4 temperature regulating device, 4.1 heat dissipation fins, 4.2 fan, 5 first circulation loop, 5.1 circulation pump, 6 second circulation pipeline, 6.1 first capillary, 6.2 second capillary, 6.3 third capillary, 6.4 peristaltic pump, 6.5 first tee, 6.6 second tee, 6.7 capillary entry end, 6.8 capillary exit end, 7 opening and closing cover, 7.1 cavity structure, 8 PCB board, 9 capillary, 10 disposable rubber head. DETAILED DESCRIPTION

[0022] In order to make the purpose, principle and structure of the present invention more clear, it is further described below in conjunction with the drawings and specific embodiments.

[0023] Embodiment 1:

[0024] This embodiment provides a capillary liquid cooling column oven, see Figures 1 to 4 include:

[0025] The box body 1 has a display screen and a touch button device on the front side thereof, and an opening and closing cover 7 connected to the box body 1 by a pin shaft. The opening and closing cover 7 and the front side of the box body 1 form a cavity structure 7.1, and the side of the opening and closing cover 7 is also provided with a plurality of openings.

[0026] A temperature regulating container 2, a liquid storage container 3 and a temperature regulating device 4 are arranged in the box body 1.

[0027] A first circulation loop 5 is provided between the temperature regulating container 2 and the liquid storage container 3. The first circulation loop 5 comprises: a plurality of pipelines sealed and connected to the temperature regulating container 2 and the liquid storage container 3 and a circulation pump 5.1 connected to the pipelines.

[0028] The front end of the liquid storage container 3 is sealed and connected to a plurality of thin tubes, and the plurality of thin tubes pass through the front panel of the box body 1 and extend into the cavity structure 7.1. The plurality of thin tubes, the plurality of tees, the liquid storage container 3 and the peristaltic pump 6.4 together form a second circulation loop 6. The second circulation loop 6 specifically includes: a first thin tube 6.1, one end of which is connected and connected to the first liquid outlet of the liquid storage container 3, and the other end is connected to the first tee 6.5; a third thin tube 6.3, one end of which is connected and connected to the first liquid inlet of the liquid storage container 3, and the other end is connected to the second tee 6.6; a second thin tube 6.2, both ends of which are connected and connected to the first tee 6.5 and the second tee 6.6 respectively.

[0029] Furthermore, the first capillary 6.1 is also connected to a peristaltic pump 6.4, and the flow rate of the peristaltic pump 6.4 is 0.5 ml / s. The peristaltic pump 6.4 allows the coolant to circulate in the capillary at a lower flow rate and a lower liquid pressure, so that the capillary 9 and the coolant in the capillary can fully exchange heat without causing a sudden change in temperature, and the sudden change in the temperature difference between the coolant and the capillary 9 is also reduced, thereby avoiding condensation in the capillary during the cooling process, and the lower liquid pressure can ensure that the sealing measures at the tee will not be broken by the liquid pressure.

[0030] The remaining end of the first tee 6.5 not connected to the capillary is a capillary entry end 6.7, and the remaining end of the second tee 6.6 not connected to the capillary is a capillary exit end 6.8. The capillary entry end 6.7 and the capillary exit end 6.8 are provided with a disposable rubber head 10, which can be pierced by the capillary 9 and can fill the gap between the capillary 9 and the tee and the capillary through elastic deformation, thereby forming a seal between the capillary 9 and the second circulation loop 6 after puncture.

[0031] The temperature regulating device 4 comprises a plurality of heat dissipating fins 4.1 and fans 4.2 which are arranged alternately.

[0032] The temperature regulating container 2 includes a main circulation pipe 2.1 and a plurality of branch circulation pipes 2.2. The main circulation pipe 2.1 is sealed and connected with the branch circulation pipes 2.2. The branch circulation pipes 2.2 pass through the fan 4.2 and the heat dissipation fins 4.1 in the temperature regulating container 2. The main circulation pipe 2.1 is also sealed and connected with the first circulation loop 5. In the temperature regulating container 2, the outside of the main circulation pipe 2.1 and the branch circulation pipes 2.2 are also wrapped with thermal insulation cotton.

[0033] A PCB board 8 is also provided in the chassis 1, and the PCB board 8 can carry a number of electronic components for signal conduction with the display screen, touch button device, circulation pump 5.1, peristaltic pump 6.4, and temperature adjustment device 4 to achieve control of the above devices.

[0034] The specific working principle is as follows:

[0035] Step 1: Turn on the device power and set the relevant operating requirements and standards on the touch button device.

[0036] Step 2, open the opening and closing cover 7, and pierce the capillary 9 through the disposable rubber heads 10 on the first tee 6.5 and the second tee 6.6 at one time. The capillary 9 enters from the capillary entry end 6.7, passes through the first tee 6.5, the second capillary 6.2 and the second tee 6.6 in sequence, and then exits from the capillary exit end 6.8.

[0037] Step 3, closing the opening and closing cover 7, at which time the capillary 9 can pass through the opening hole on the opening and closing cover 7 without forming a limit with the opening and closing cover 7. The isolation design of the opening and closing cover 7 avoids the room temperature from affecting the cooling of the capillary 9.

[0038] Step 4, operate the touch key device start device, and the circulation pump 5.1 and the peristaltic pump 6.4 are started in sequence.

[0039] Step 4.1, when the circulation pump 5.1 is started, the fan 4.2 is started, and the coolant in the branch circulation pipes 2.2 passing through the temperature regulating device 4 is cooled by the fan 4.2 and the heat dissipation fins 4.1, and the coolant in the branch circulation pipes 2.2 and the main circulation pipe 2.1 is cooled together by thermal convection.

[0040] Step 4.2, the circulation pump 5.1 extracts and circulates the coolant in the temperature regulating container 2 and the coolant in the liquid storage container 3, so that the coolant in the temperature regulating container 2 that has been cooled enters the liquid storage container 3 and forms a buffer and temporary storage.

[0041] In step 4.3, the peristaltic pump 6.4 pumps the coolant in the liquid storage container 3 into the first circulation loop 6. Under the negative pressure extraction of the peristaltic pump 6.4, the coolant in the liquid storage container 3 passes through the first capillary 6.1, the second capillary 6.2 and the third capillary 6.3 in sequence. Among them, the capillary 9 passes through the second capillary 6.2 and its outer wall is wrapped by the coolant in the second capillary 6.2, realizing heat exchange between the coolant and the capillary 9, and taking away the Joule heat generated during the electrophoresis process.

[0042] Step 5: After the electrophoresis operation is completed, continue to circulate the coolant for a period of time until the coolant temperature and the capillary temperature tend to be stable. This measure avoids condensation in the capillary due to unnecessary temperature changes.

[0043] Step 6, pull out the capillary 9, close the tee, replace the disposable rubber head 10, and shut down the machine.

[0044] Embodiment 2:

[0045] In this embodiment, referring to the "Capillary Electrophoresis JJG 964-2001" of the "National Metrology Verification Regulations of the People's Republic of China", a standard test was performed on vitamin B6 (abbreviated as "VB6"), and a comparative test was performed under two conditions: using the capillary liquid-cooled column oven of the present invention and not using the capillary liquid-cooled column oven of the present invention.

[0046] The experiments all used a CE-1000 capillary electrophoresis instrument, a wavelength environment of 254 nm, a 75 um inner diameter capillary, a sample containing 0.1 mg / mL VB6 (AUX=1), a high-voltage power supply operating voltage of 15 kV, a current variation of 34 uA~38 uA, an injection time of 3 s, an injection pressure of 8 kPa, and a 20 mM disodium hydrogen phosphate electrophoresis buffer.

[0047] Experimental group 1: The capillary liquid-cooled column oven of the present invention was not used.

[0048] The test data of 3-pin VB6 sample CE-UV at room temperature (20°C) are as follows:

[0049] ;

[0050] Table 1: Test data table without using the capillary liquid cooling column oven of the present invention.

[0051] The test results are shown in the above data. After injection at room temperature, the repeatability RSD (%) of 3 injections of VB6 samples is 16.35% for qualitative repeatability and 40.2% for quantitative repeatability, which does not meet the requirements of the national metrological verification regulations. The test data is invalid. The chromatogram can be found in Figure 5 .

[0052] Experimental group 2: using the capillary liquid-cooled column oven of the present invention.

[0053] The test data of 3-pin VB6 sample CE-UV under 4℃ condition is as follows:

[0054] ;

[0055] Table 2: Test data table of the capillary liquid-cooled column oven of the present invention.

[0056] The test results are shown in the data section above. After using the capillary liquid-cooled column oven, the repeatability RSD (%) of the 3-pin VB6 sample, the qualitative repeatability is less than 1%, the quantitative repeatability is less than 1%, the theoretical plate number is maintained above 13,000, and the column efficiency is optimal, fully meeting and exceeding the requirements of the national metrological verification regulations. The chromatogram can be found in Figure 6 .

[0057] Furthermore, combining the above test data and Figure 5 , Figure 6By comparing and analyzing the chromatograms, we can conclude that when injected at room temperature, the repeatability of the VB6 sample is extremely poor, and both the qualitative and quantitative repeatability are far from meeting the requirements of the national metrological verification procedures, and the monitoring data is invalid. After adding the use of a capillary liquid-cooled column oven, the qualitative and quantitative repeatability RSD% of the three-injection VB6 samples were less than 1%, the theoretical plate number exceeded 13,000, and the column efficiency reached the best and far exceeded the standard. This fully demonstrates that the capillary liquid-cooled column oven can effectively stabilize the experimental conditions, significantly improve the accuracy and reliability of the capillary electrophoresis results, and is a key equipment to ensure the accuracy and compliance of experimental data, and plays an important role in improving the quality of capillary electrophoresis detection.

[0058] Embodiment three:

[0059] This embodiment provides a capillary electrophoresis analyzer, which uses a capillary liquid-cooled column oven as described in the first embodiment.

[0060] The electrophoresis analyzer comprises: a capillary, both ends of which are immersed in a buffer solution; a sample injection head arranged on the capillary; an electrophoresis zone arranged on the capillary and on the right side of the sample injection head; a detector arranged on the capillary and on the right side of the electrophoresis zone; two buffer tanks arranged on both ends of the capillary and used to contain buffer solutions; and a high-voltage electrical system whose both ends are electrically connected to the buffer solutions in the two buffer tanks. The portion of the capillary between the sample injection head and the detector is arranged in the second capillary of the capillary liquid-cooled column oven described in the first embodiment.

[0061] The above description is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent substitutions or changes according to the technical solutions and novel concepts of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A capillary liquid-cooled column oven, comprising a box body, characterized in that: Also includes: The temperature regulating container and liquid storage container are arranged in the box. A temperature regulating device is provided on one side of the temperature regulating container to fit the temperature regulating container, and the temperature regulating container forms a first circulation loop with the liquid storage container through a circulation pump and a circulation pipeline; The liquid storage container forms a second circulation loop through a capillary tube and a peristaltic pump. The second circulation loop comprises a first capillary tube, a second capillary tube and a third capillary tube, one end of the first capillary tube is connected to the first liquid outlet of the liquid storage container, one end of the third capillary tube is connected to the first liquid inlet of the liquid storage container, and the other ends of the first capillary tube and the third capillary tube are respectively connected to the two ends of the second capillary tube through a tee; The tee is used to puncture the capillary, so that the capillary enters from one end of the first tee, passes through the second capillary, and then exits from one end of the second tee; The temperature regulating container includes a main circulation pipe and several metal branch circulation pipes. The branch circulation pipes are closed and connected with the main circulation pipe after passing through the temperature regulating device. The main circulation pipe is also closed and connected with the first circulation loop. The temperature regulating container is wrapped with insulation material on the outside of the main circulation pipe and the branch circulation pipe.

2. A capillary liquid-cooled column oven as claimed in claim 1, characterized in that: An opening and closing cover is provided on the front side of the box body, one end of which is pin-connected to the front side of the box body, and the opening and closing cover and the front side of the box body form a cavity structure. The capillary tube of the second circulation loop passes through the through hole on the front side of the box body and is arranged in the cavity structure.

3. A capillary liquid-cooled column oven as claimed in claim 1, characterized in that: The temperature regulating device comprises a plurality of heat dissipation fins and a plurality of fans, and the heat dissipation fins and the fans are arranged alternately.

4. A capillary liquid-cooled column oven as claimed in claim 3, characterized in that: A PCB board is also provided in the box body, and the PCB board is connected with the temperature regulating device, the circulation pump of the first circulation loop and the peristaltic pump of the second circulation loop by signal to form signal control.

5. A capillary liquid-cooled column oven as claimed in claim 1, characterized in that: The capillary entry end of the first tee and the capillary exit end of the second tee are provided with disposable puncturing rubber heads, so that the capillary punctures through and forms a liquid seal between the capillary and the tee through elastic deformation.

6. A capillary liquid-cooled column oven as claimed in claim 1, characterized in that: The flow rate of the peristaltic pump is 0.5 ml / s.

7. A capillary electrophoresis analyzer, comprising: A capillary tube, both ends of which are immersed in a buffer solution respectively; A sample injection head disposed on the capillary; An electrophoresis zone set on the capillary and on the right side of the sample injection head; A detector is arranged on the capillary and on the right side of the electrophoresis zone; two buffer tanks are arranged at both ends of the capillary and are used to contain buffer; a high-voltage electrical system whose two ends are electrically conductive with the buffer in the two buffer tanks, characterized in that the part of the capillary between the sample injection head and the detector is arranged in the second capillary liquid-cooled column oven as described in any one of claims 1-6.

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